WO1999026642A1 - Use of the e.coli strain dsm 6601 for treating diarrhoea in veterinary medicine - Google Patents

Abstract

The invention relates to the use of the Escherichia coli strain DSM 6601 to produce medicaments for preventing and treating microbially-caused diarrhoea involving pathogenic fungi in animals.

Description

Use of Escherichia coli strain DSM 6601 for the treatment of diarrhea in the veterinary sector

The invention relates to the use of Escherichia coli strain DSM 6601 for the prophylaxis and treatment of diarrhea caused by robots in animals.

Diarrhea in humans or animals means the frequent emptying of thin-bodied faeces, the cause of which can vary, for example an allergic reaction to certain foods, but indicates a reaction to microorganisms, namely bacteria, fungi or viruses or their toxins. Prolonged diarrhea can lead to serious complications up to the exitus due to the strong loss of water and salt. Severe diarrhea is therefore definitely in need of treatment, especially if it concerns young or already weakened people or animals.

In a majority of cases, diarrhea is caused by bacteria that do not belong to the normal microorganism flora of the body in question, with particular pathogenic strains of Escherichia coli, Salmonella and Shigella playing a major role. In addition to bacteria, diarrhea can also be triggered by infections with viruses, especially corona and rotaviruses, or by fungi, which in most cases are classified as Eumykota. While bacterial diarrhea can usually be fought quite well today by the administration of sorbents or non-systemic sulfonamides or antibiotics, there are still no drugs that are successful in viral infections can be used and hardly any drugs that are effective in colonizing the gastrointestinal tract with fungi.

Fungi are a polyphylogenetic group of chlorophyll-free, heterotrophic and eukaryotic organisms that can be single or multicellular and whose multiplication, like with all eukaryotes, occurs with mitosis and meiosis and through the formation of sexual or asexual spores, but also through sprouts. The unicellular mushrooms, which only reproduce through sprouts, are often grouped together as yeasts, although the yeasts are a conglomerate of different families. Almost all Eumykota, i.e. the real mushrooms, go through two or more morphologically distinguishable stages during individual development, namely as teleomorphs, in which the spores are formed after meiosis, and one or more stages as anamorphs, in which the spore formation is not in connection with the Meiosis stands. Fungi whose teleomorphs are unknown or which have lost the ability to form them are grouped together as fungi imperfecti. The exact botanical classification of the mushrooms is often still open.

Due to their lack of chlorophyll, all fungi feed heterotrophically through the breakdown of organic substances; so they are saprophytes. Medically, a distinction is made between the opportunists, i.e. saprophytes, which can become pathogenic rarely and only under very specific conditions and which include, for example, Candida and Aspergillus. So-called pathogenic saprophytes lead a normal saprophytic life exogenously, but they are to be considered pathogenic for humans or animals in all circumstances if an infection has occurred Has. There are also obligatory parasites among the fungi, which, moreover, cannot vegetate the host organism and which are only found in humans or animals; this includes most dermatophytes.

An essential difference between bacteria and fungi lies in the fact that fungi, unlike bacteria, are eukaryotes, that is to say they have a core with a membrane and, in contrast to bacteria, also have mitochondria and that the cell wall consists of chitin and / or cellulose, while the cell wall of the bacteria is made up of mucopeptides. This also explains why most antibiotics effective in bacteria fail in fungi, because in many known antibiotics the bacterial wall or the mitochondria are the main points of attack for these substances.

There are also some antibiotics that can also be used systemically, ie not only topically, against fungal infections and that are characterized by a polyene structure, such as, for example, amphotericin B, griseofulvine, natamycin and nystatin. Synthetic systemic antimycotics are flucytosine and a number of azole derivatives such as ketoconazole and miconazole or fluconazole and itraconazole.

While the exact mechanism of action of the polyene antibiotics is not yet known in all details, the synthetic azole derivatives act on ergostorol synthesis in the cell membrane and thus influence the permeability of the cell wall. A disadvantage of all previously known antimycotics, which can also be used systemically, is that they generally do not have a fungicidal action, but only a fungistatic action, and therefore require a longer treatment period. Furthermore, there are cross-resistances in both the polyene antibiotics and in the azole compounds, and finally there is the fact that the price of these products is relatively high, which is contrary to widespread use in veterinary medicine.

There is therefore still a further need for veterinary medicinal products which are able to effectively combat diarrhea in animals which are essentially or also due to the involvement of pathogenic fungi.

Completely surprisingly, it has now been found that when using Escherichia coli strain DSM 6601 such diarrhea can be combated well, even if the usual medication with fungus-active compounds has failed.

Escherichia coli, hereinafter abbreviated as E. coli, occurs in numerous variants which differ in terms of capsule antigens, surface antigens and flagella antigens and can therefore be divided into numerous serological types. The classification according to serotypes does not mean anything about the different virulence of the pathogens. Representatives of one and the same serotype can have a different pathogenicity potential in both the human and animal body, which in extreme cases can range from avirulent to highly pathogenic. However, it is known that the E. coli strain DSM 6601 is not human or animal pathogen is assessed. This strain is used, for example, in human medicine as a substitute preparation for infectious bowel diseases caused by Salmonella or Shigella, in both acute and chronic cases. This E. coli strain is also successfully used in substitution therapy for other disorders of the intestinal flora, such as after antibiotic treatment or radiation. Whether this is actually just the displacement of the pathogenic bacterial strains with the corresponding variants of E. coli or Proteus by this special E. coli strain, i.e. a reduction in the toxins, or whether the metabolic products of the E. coli DSM strain Developing a therapeutic effect on its own has not been clearly clarified.

On the basis of these known explanations of the mechanism of action of living E. coli germs, however, it was in no way to be expected that treatment with these living cultures would show a surprisingly extensive effectiveness in infecting the intestinal tract of animals with fungi and here in particular yeasts . In the 1950s, apathogenic E. coli strains had occasionally been used in veterinary medicine for diseases of cattle or pigs, some of which were also associated with diarrhea, but this was the treatment of nutritional disorders in piglets (Fischer W ., Experience of a practical veterinarian in the treatment of sick piglets from 1945 to 1950; dissertation, University of Munich 1950) or in the treatment of so-called Semper disease in cattle, which according to the results of this publication was obviously based on malnutrition due to the geology of the area ( Häfele W., The "Semper Disease" a nutritional and developmental disorder of the cattle in the Black Forest in the Surroundings of St. Blasien; Dissertation, vet. med. Veterinary clinic, University of Munich 1952). These early attempts to use certain strains of coli on a trial basis for special diseases of pigs or cattle were completely isolated and did not even give rise to further attempts of this type in the case of bacterial diarrhea. It was all the more surprising to find that the E. coli strain DSM 6601 shows a surprising effect even in intestinal diseases that are exclusively or essentially caused by fungi, since the fight against fungal infections, especially when the intestinal mucous membranes are affected, is particularly difficult since the preparations used for bacterial infections have practically no effect.

To date, surprisingly little is known about the normal intestinal flora in various animal species, but an infestation of the gastrointestinal tract with fungi and especially with yeasts can always be interpreted as a pathological event.

The surprisingly rapid effectiveness of the treatment with E. coli strain DSM 6601 leads to the conclusion that the effect of this treatment is not based or not only on the substitution of the fungal flora by a healthy bacterial flora, but that the strain greatly increases the body's own Defense contributes, whereby it can be assumed that the metabolic products of this strain have a considerable immunostimulatory effect.

The invention is explained in more detail below using an example: Example:

In a dairy farm in Saxony, which had an average of 1,480 dairy cows, 600 heifers and young cattle and 185 calves, the calf population was characterized by very good rearing results, low morbidity and mortality until early January 1997. Manifest gastric and intestinal diseases with diarrhea were extremely rare. Calf losses were always below the 3% limit based on the number of live calves.

At the end of January 1997, diarrhea suddenly occurred in the herd with clinically striking symptoms of gastroenteritis. The disease began in the calves between the ages of seven and eleven days, was of varying duration and was characterized by a morbidity rate of more than 90% and a mortality rate of more than 10%. The course of the disease was characterized by the following symptoms: initially greenish-yellowish diarrhea with a thick pulpy consistency, then the feces became slimy, increasingly thinly pulpy and mostly watery as the disease progressed. The body temperature values were slightly elevated at the beginning of the disease (39.7 to 39.9 ° C), but then fell rapidly in the further course and only reached lower limits, sometimes in the range of 37.8 to 37.5 ° C. The sick calves showed pronounced reluctance to suckle and increased weakness, so that it became stuck in the further course of the disease. Critically ill animals had to be force-fed.

After a period of three to nine days, the losses increased. The sections carried out showed agreement highly striking findings, namely a fluid, fluffy to slightly bloody intestinal contents and high-grade, ulcerative stomach and abdominal inflammation. In April 1997, the incidence of disease reached nearly 100% and the mortality rose to more than 15%.

The microbiological studies of the contents of the stomach and intestines and of the internal organs were initially without any specific results, until in February 1997 yeasts of the genus Candida (C. glabrata and C. albicans) were isolated from two stomach calves from stomach and intestine contents. Intestinal pathogens E. coli were not found. These yeasts were detected in all other sections of the dead calves and in the examination of faecal samples from calves and their mothers. Further targeted examinations of faeces and colostrum samples from the cows and heifers brought the same results. Additional examinations for viruses, namely corona and rotaviruses, as well as for cryptosporidia, revealed coronavirus particles only in two cases when calves died and rotaviruses in one case.

The origin of the yeasts could not be explained for a long time until in March 1997 when investigating beer spent grains used as animal feed, it was possible to detect yeasts (dominating Candida glabrata) in an amount of 3.6 x 10 ⁷ CFU / g animal feed.

The therapeutic measures that were carried out extensively right from the start, such as diet drinks, anti-inflammatory drugs, styptics, electrolytics and cardiovascular agents, only led to unsatisfactory results. When using The animals died of sulfonamides and antibiotics six to eight hours after application. After these therapies were unsuccessful, therapeutic agents containing humic acids as effective substances were used. These measures also did not result in any noticeable improvements in the course of the disease.

Of the 236 calves born alive in two months from April 24 to June 23, 1997, all animals fell ill, and 41 animals died during this period, which corresponds to 1 7.4%, 29 animals due to diarrhea corresponding to 70, 7% of the losses and eight animals in diarrhea with accompanying bronchopneumonia corresponding to 19.5% of the losses and four animals for other reasons, corresponding to 9.8% of the losses.

In the period from June 24, 1997 to September 4, 1997, 300 newborn calves were given a suspension of live E. coli strain DSM 6601 in an amount of 15.0 ml per calf per day corresponding to ... CFU / ml. This dose was independent of the weight and age of the animal and was administered orally, namely over a period of 10 to 13 days post partum. During the period from June 24 to July 8, 1,997, 61 calves were treated in this way, two of whom developed diarrhea, which corresponds to a morbidity of 3.3%. None of the animals died from this disease.

Between July 9 and July 23, 1997, 49 calves were treated accordingly, none of which suffered from gastroenteritis with diarrhea. The mortality from this disease was 0%. During the period from July 23 to August 6, 1997, 64 calves were treated, two of which suffered from diarrhea, which corresponds to a morbidity of 3.1%. None of the animals died from this disease, so that the mortality is 0.

After all available therapeutic options had been exhausted, the prophylactic and therapeutic use of an E. coli strain DSM 6601 suspension almost completely prevented the occurrence of diarrhea due to gastroenteritis in suckling calves.

The use of other medicines and dietetics could be reduced considerably, so that the costs for this were reduced by 70%.

Claims

claims

1 . Use of Escherichia coli strain DSM 6601 for the production of pharmaceuticals for the prophylaxis and treatment of microbial! caused diarrhea with the participation of pathogenic fungi

Animals.

BBATZBLÄTT (LEVEL 26)